Abstract：Macropsinae is widely distributed around the world. The phylogenetic relationship of the species in Macropsinae appears to be confused, and the taxonomic status of some genera strongly needs to be verified and discussed. 23 Cicadellidae species (including 22 Macropsinae of ingroup and 1 Idiocerinae of outgroup) were chosen as the materials and 23 DNA sequences of mitochondrial COⅠ gene were obtained. The sequence composition and genetic distance of these genes were analyzed, and the phylogenetic trees were built using the methods of Maximum Likelyhood and Bayesian Inference. The results showed that the content of A+T (68.3%) in the sequences of COⅠ gene was significantly higher than that of G+C (31.7%). There were statistically significant differences in the genetic distance among species, genera and subfamilies. The phylogenetic tree well supported the monophyly of genus Oncopsis and Pedionis, but weakly supported that of Macropsis and Pediopsoides.